Bordetella pertussis is the bacterial pathogen responsible for whooping cough in humans. Presently, the only widely-available commercial diagnostic probe available to detect B. pertussis in naso-pharyngeal aspirates or in cultures from patients with clinical symptoms of pertussis is a crude polyclonal antiserum that typically gives high background in routine immunofluorescence assays.
In addition to its diagnostic applications, polyclonal antiserum to B. pertussis has also been employed to establish serotypes for B. pertussis. Serotype markers for the bacterium have been defined by the ability of strain-specific polyclonal antisera to agglutinate the bacteria.
E. K. Andersen first identified five distinctive agglutinogen factors in 1953 (Acta Pathol. Microbiol. Scand. 33:202-224 (1953)), and Eldering et al. subsequently added agglutinogen factor 6 (J. Bacteriol. 74:133-136 (1957)). 1n fact, it is the use of mono-specific polyclonal antisera (Preston factors 1-3) in combination with U.S. Reference Factor 1-6 antisera which has permitted B. pertussis to be distinguished from the closely related species B. bronchiseptica and B. parapertussis.
Following whooping cough outbreaks, it has been noted that there tends to be a prevalence of certain B. pertussis serotypes, and the serum agglutinin titers of human vaccinees appear to correlate with clinical protection from pertussis. Thus, identification of specific agglutinogens to B. pertussis is of potential importance since they may serve as protective antigens.
Some of the agglutinogen factors have been defined, for instance, the expression of lipooligosacccharide A (LOS A) by B. pertussis cells appears to correlate with the presence of the serotype 1 agglutinogen factor. Likewise, fimbrial agglutinogens have been found to correspond to serotype 2-, 3-and 6-containing cells, and a nonfimbrial 69 kDa outer membrane protein appears to correlate to serotype 3 cells.
In view of the above, the present inventors sought to create Bordetella pertussis hybridomas and serotype-specific monoclonal antibodies (Mabs) therefrom in order to specifically detect B. pertussis organisms and the antigens that they produce, and to advantageously utilize such Mabs for diagnostic, manufacturing and research purposes.